RNA-seq转录组测序分析不同季节对槟榔江水牛精液品质的影响

doi:10.11843/j.issn.0366-6964.2016.07.009

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (7): 1373-1380.doi: 10.11843/j.issn.0366-6964.2016.07.009

RNA-seq转录组测序分析不同季节对槟榔江水牛精液品质的影响

耿红红，张敬旸，李莲，王根林^*

（南京农业大学动物科技学院，南京 210095）

收稿日期:2016-03-22 出版日期:2016-07-23 发布日期:2016-07-23
通讯作者: 王根林，教授，E-mail:genlwang@sina.com
作者简介:耿红红(1989-)，女，山西孝义人，硕士生，主要从事动物生殖与生理调控研究，E-mail：15150537075@163.com
基金资助:
国家自然科学基金项目（31372290）;“十二五”国家科技计划项目（2012BAD12B10）

Semen Quality Analysis of Betelnut-jiang Buffalo in Different Seasons by RNA-seq

GENG Hong-hong，ZHANG Jing-yang，LI Lian，WANG Gen-lin^*

(College of Animal Science and Technology，Nanjing Agricultural University，Nanjing 210095，China)

Received:2016-03-22 Online:2016-07-23 Published:2016-07-23

摘要/Abstract

摘要：

本研究旨在利用RNA-seq技术对不同季节槟榔江水牛全血mRNA进行分析，找出不同季节精液品质候选基因。选取5头生长状况相近的3~5岁槟榔江水牛，进行春、夏、秋、冬4个季节的精液品质（鲜精活力、精子畸形率、射精量、精子密度）检测。根据精液品质，选取差异极显著的春（BL）、夏（SBL）两个季节的水牛样品进行RNA-seq转录组测序分析。颈静脉采集全血，抽提mRNA并组内混合制样，利用RNA-seq技术分析不同季节槟榔江水牛全血mRNA差异表达。根据差异倍数>1.5，P<0.05的标准进行筛选，得到差异基因，再利用DAVID 6.7软件对差异基因进行GO和KEGG分析，通过RT-PCR验证基因芯片结果的准确性。结果表明：差异表达基因84个，其中上调基因56个，下调基因28个；GO分析发现，这些差异基因与应激反应、免疫反应有关；KEGG分析表明，差异基因显著富集到肿瘤坏死因子（TNF）和T细胞受体信号通路。其中，神经营养因子受体类似物死亡结构域（NRADD，又叫热休克蛋白因子1 (HSF1)）、原癌基因FOS和JUN，与精液品质相关性强，可作为进一步研究的靶基因。

Abstract:

To determine the key candidate genes related to semen quality in different seasons，the mRNA profiling of Chinese Betelnut-jiang buffalo were analyzed by RNA-seq.Based on similarities in growth status，5 Betelnut-jiang buffalo bulls were selected and their semen quality were tested (including ejaculate volume，fresh sperm motility，sperm density and abnormality rate).According to the semen quality，the samples of summer (treated as SBL group) and spring (as BL group) were selected for RNA-seq.We collected the whole blood in jugular vein of Betelnut-jiang buffalo bulls，extracted the mRNAs and mixed them within group.Then，the mRNA profiling was used to measure the mRNAs in Betelnut-jiang buffalo bulls.Gene expression levels with >1.5 fold change and P value < 0.05 between the BL and SBL groups were further analyzed.Furthermore，GO and KEGG were used to analyze the genes enrichment.Finally，RT-PCR was used to confirm the RNA-seq result.The results indicated 84 differentially expressed genes.Among these，56 genes were up-regulated and 28 genes were down-regulated in SBL group.The GO search revealed that these differential expression genes were mainly related to stress reaction and immune response.And the KEGG selected the TNF signaling pathway and T cell receptor signaling pathway.Neurotrophin receptor alike death domain（NRADD，also named heat shock factor protein 1(HSF1)），localisation of FOS (FOS) and jun proto-oncogene (JUN)，were strongly associated with semen quality in different seasons which could be used as target genes for further study.

中图分类号:

S823.8+3

耿红红，张敬旸，李莲，王根林. RNA-seq转录组测序分析不同季节对槟榔江水牛精液品质的影响[J]. 畜牧兽医学报, 2016, 47(7): 1373-1380.

GENG Hong-hong，ZHANG Jing-yang，LI Lian，WANG Gen-lin. Semen Quality Analysis of Betelnut-jiang Buffalo in Different Seasons by RNA-seq[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1373-1380.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2016.07.009

https://www.xmsyxb.com/CN/Y2016/V47/I7/1373

参考文献

［1］HAMILTON T R，MENDES C M，DE CASTRO L S，et al.Evaluation of lasting effects of heat stress on sperm profile and oxidative status of ram semen and epididymal sperm［J］.Oxid Med Cell Longev，2016，2016：1687657.
［2］KOIVISTO M B，COSTA M T，PERRI S H，et al.The effect of season on semen characteristics and freezability in Bos indicus and Bos taurus bulls in the southeastern region of Brazil［J］.Reprod Domest Anim，2009，44(4)：587-592.
［3］HELBIG L，WOODBURY M R，HAIGH J C，et al.The seasonal fertility of North Ameriman bison (Bison bison) bulls［J］.Anim Reprod Sci，2007，97(3-4)：265-277.
［4］ROCHA A，RANDEL R D，BROUSSARD J R，et al.High environmental temperature and humidity decrease oocyte quality in Bos taurus but not in Bos indicus cows［J］. Theriogenology，1998，49(3)：657-665.
［5］王宝全，孔伟，马志远，等.高温对种公牛精液品质的影响研究［J］.中国牛业科学，2013，39(5)：13-15.
WANG B Q，KONG W，MA Z Y，et al.Influence of high temperature on semen quality［J］.China Cattle Science，2013，39(5)：13-15.(in Chinese)
［6］MAYAHI S，MAMOUEI M，TABATABAEI S，et al.Reproductive characteristics and thyroidal function in relation with season in Khuzestan buffalo (Bubalus bubalis) bulls［J］.Vet Res Forum，2014，5(3)：201-205.
［7］刘红亮，郑丽明，刘青青，等.非模式生物转录组研究［J］.遗传，2013，35(8)：955-970.
LIU H L，ZHENG L M，LIU Q Q，et al.Studies on the transcriptomes of non-model organisms［J］.Hereditas，2013，35(8):955-970.(in Chinese)
［8］YAN L，YANG M，GUO H，et al.Single-cell RNA-Seq profiling of human preimplantation embryos and embryonic stem cells［J］.Nat Struct Mol Biol，2013，20(9)：1131-1139.
［9］贾昌路，张瑶，朱玲，等.转录组测序技术在生物测序中的应用研究进展［J］.分子植物育种，2015，13(10)：2388-2394.
JIA C L，ZHANG Y，ZHU L，et al.Application progress of transcriptome sequencing technology in biological sequencing［J］. Molecular Plant Breeding，2015，13(10)：2388-2394.(in Chinese)
［10］张汝，赵家才，毛翔光，等.槟榔江水牛精液品质的研究［J］.云南畜牧兽医，2013(5)：20-22.
ZHANG R，ZHAO J C，MAO X G，et al.The semen quality study of Bos bubalus bulls［J］.Yunnan Animal Husbandry & Veterinary Medicine，2013(5)：20-22.(in Chinese)
［11］桑润滋，田树军，李俊杰，等.热应激对荷斯坦种公牛精液品质的影响及机理［J］.中国兽医学报，2002，22(4)：407-410.
SANG R Z，TIAN S J，LI J J，et al.Influence of heat stress on semen quality of Holstein bull and approach to its mechanism［J］.Chinese Journal of Veterinary Science，2002，22(4)：407-410.(in Chinese)
［12］DURAIRAJANAYAGAM D，AGARWAL A，ONG C.Causes，effects and molecular mechanisms of testicular heat stress［J］.Reprod Biomed Online，2015，30(1)：14-27.
［13］AKERFELT M，MORIMOTO R I，SISTONEN L.Heat shock factors：integrators of cell stress，development and lifespan［J］.Nat Rev Mol Cell Biol，2010，11(8)：545-555.
［14］XU D，SUN L，LIU S，et al.Molecular cloning of hsfll and hsbpl cDNAs，and the expression of hsfl，hsbpl and hsp70 under heat stress in the sea cucumber Apostichopus japonicus［J］.Comp Biochem Physiol B Biochem Mol Biol，2016，198：1-9.
［15］ABANE R，MEZGER V.Roles of heat shock factors in gametogenesis and development［J］.FEBS J，2010，277(20)：4150-4172.
［16］HAYASHIDA N，INOUYE S，FUJIMOTO M，et al.A novel HSF1-mediated death pathway that is suppressed by heat shock proteins［J］.EMBO J，2006，25(20)：4773-4783.
［17］NAKAI A，SUZUKI M，TANABE M.Arrest of spermatogenesis in mice expressing an active heat shock transcription factor 1［J］.EMBO J，2000，19(7)：1545-1554.
［18］KORFANTY J，STOKOWY T，WIDLAK P，et al.Crosstalk between HSF1 and HSF2 during the heat shock response in mouse testes［J］.Int J Biochem Cell Biol，2014，57：76-83.
［19］ROBERTSON K M，O’DONNELL L，JONES M E，et al.Impairment of spermatogenesis in mice lacking a functional aromatase (cyp 19) gene［J］.Proc Natl Acad Sci U S A，1999，96(14)：7986-7991.
［20］况海斌，方廉.原癌基因与睾丸功能的关系［J］.中华男科学，2003，9(5)：377-380.
KUANG H B，FANG L.The relationship between proto-oncogene and testicular function［J］.National Journal of Andrology，2003，9 (5)：377-380.(in Chinese)
［21］WESTON C R，DAVIS R J.The JNK signal transduction pathway［J］.Curr Opin Cell Biol，2007，19(2)：142-149.
［22］HALL S H，BERTHELON M C，AVALLET O，et al.Regulation of c-fos，c-jun，jun-B，and c-myc messenger ribonucleic acids by gonadotropin and growth factors in cultured pig Leydig cell［J］.Endocrinology，1991，129(3)：1243-1249.
［23］杜琛，付绍印，高鸿雁，等.绒山羊肌内脂肪细胞成熟前后比较转录组分析［J］.畜牧兽医学报，2014，45(5)：714-721.
DU C，FU S Y，GAO H Y，et al.Transcriptome analysis of intramuscular preadipocytes and mature adipocyte in cashmere goats［J］. Acta Veterinaria et Zootechinca Sinica，2014，45(5)：714-721.(in Chinese)

[1]	陈凌声，代小丽，徐永茹，罗婵，陆凤花，蒋建荣，石德顺，徐平，李湘萍. 成熟前后水牛卵母细胞蛋白质组的初步研究[J]. 畜牧兽医学报, 2016, 47(10): 2003-2011.
[2]	苏节，刘庆友，朱鹏，沈开元，石德顺，崔奎青. 沼泽型水牛CYP19A1基因克隆、序列分析及组织表达研究[J]. 畜牧兽医学报, 2013, 44(4): 514-521.

RNA-seq转录组测序分析不同季节对槟榔江水牛精液品质的影响

Semen Quality Analysis of Betelnut-jiang Buffalo in Different Seasons by RNA-seq

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价